Cromatest HCV Test Device

PRINCIPLE

Linear Anti-HCV cassette detects antibodies (IgG, IgM, IgA) to HCV through visual interpretation of color development in the internal strip. Protein A is immobilized on the test region of the membrane. During testing, the specimen reacts with recombinant HCV antigen conjugated to colored particles and pre-coated onto the sample pad of the test. The mixture then migrates through the membrane by capillary action and interacts with reagents on the membrane. If there are sufficient HCV antibodies in the specimen, a colored band will form at the test region (T) of the membrane. The presence of this colored band indicates a positive result, while its absence indicates a negative result. The appearance of a colored band at the control region (C) serves as a procedural control, indicating that the proper volume of specimen has been added and membrane wicking has occurred. The (T) line is pre- coated with recombinant HCV fusion antigen (core, NS3, NS4 and NS5), and the C line is pre-coated with a control line antibody

REAGENT COMPOSITION

HCV test devices, contains protein A coated particles and HCV antigen coated on the membrane.

STORAGE AND STABILITY

Store at 2-30ºC. The test cassette is stable through the expiration date printed on the sealed pouch. The cassette must remain in the sealed pouch until use. DO NOT FREEZE. Do not use beyond the expiration date or devices with damaged pouch. Do not reuse tests. Care should be taken to protect the components of the kit from contamination. Do not use if there is evidence of microbial contamination or precipitation. Biological contamination of dispensing equipment’s or reagents can lead to false results.

SPECIMEN COLLECTION AND PREPARATION

• Serum and Plasma. Collect blood aseptically by venipuncture into plain, heparinized or EDTA tubes, and separate the serum or plasma from the cells by centrifugation. Separate serum or plasma from the blood as soon as possible to avoid hemolysis. Test samples as soon as possible after collecting. Store samples at 2-8°C if not tested immediately. Specimens might be stored at 2°- 8°C for up to 5 days For longer storage the specimens should be frozen at -20°C. Avoid multiple freeze-thaw cycles. Prior to testing, bring frozen specimens to room temperature slowly and mix gently. Specimens containing visible particulate matter should be clarified by centrifugation before testing.Blood. Drops of whole blood can be obtained by either finge puncture or heel stick. These blood samples must be used immediately before clotting occurs. Whole blood samples containing anticoagulants should be stored in refrigeration (2-8°C) or on ice prio to testing. Whole blood samples must be tested within 24 hours o collection. Do not use any hemolyzed blood for testing

MATERIAL REQUIRED

• Specimen collection
• General laboratory
• Timer

PROCEDURE

Allow test device, specimen, buffer and/or controls to equilibrate to room temperature (15-30°C) prior to testing.

1. Remove the test device from the foil pouch and use it as soon as possible. Best results will be obtained if the assay is performed within one
2. Place the test device on a clean and level
3. Fill the plastic dropper with the specimen. Holding the plastic dropper vertically, dispense 1 drop (about 30-45 µL) of serum/plasma or 1 drop of whole blood (about 40-50 µL) into the sample well (S).

Avoid trapping air bubbles in the specimen well (S), and do not add any solution to the result area.

Immediately add 1 drop (about 35-50 µL) of Buffer into the sample well.

Avoid trapping air bubbles in the specimen well (S), and do not add any solution to the result window.

4. As the test begins to work, color will migrate across the
5. Wait for the colored band(s) to appear. The result should be read at 15 minutes.

Do not read result after 20 minutes. To avoid confusion, discard the test device after interpreting the result.

INTERPRETATION OF RESULTS

POSITIVE: *

Two colored bands appear on the membrane. One band appears in the control region (C) and another band appears in the test region (T).

NEGATIVE:

Only one colored band appears, in the control region (C). No apparent colored band appears in the test region (T).

INVALID:

Control band fails to appear. Results from any test which has not produced a control band at the specified read time must be discarded. Please review the procedure and repeat with a new test. If the problem persists, discontinue using the kit immediately.

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QUALITY CONTROL

An internal procedural control is included in the test. A colored line appearing in the control region (C) is considered an internal positive procedural control. It confirms sufficient specimen volume and correct procedural technique.

External controls are not supplied with this kit. It is recommended that positive and negative controls should be tested as a good laboratory practice to confirm the test procedure and to verify proper test performance. Handle the negative and positive controls in the same manner as patient specimens.

CLINICAL SIGNIFICANCE5

Hepatitis C Virus (HCV) is a small, enveloped, positive-sense, single- stranded RNA virus. HCV is now known to be the major cause of parenterally transmitted non-A, non-B hepatitis. Antibodies to HCV are found in over 80% of patients with well-documented non-A, non-B hepatitis. Conventional methods fail to isolate the virus in cell culture or visualize it by electron microscope.

Cloning the viral genome has made it possible to develop serologic assays that use recombinant antigens. Compared to first generation HCV EIAs using single recombinant antigens, new serologic tests include multiple antigens using recombinant protein and/or synthetic peptides to avoid nonspecific cross-reactivity and to increase sensitivity.

ANALYTICAL PERFORMANCE

Relative Sensitivity: 99%

Relative Specificity: 99.5%

Overall Agreement: 99.3%

PRECAUTIONS

1. 1. 1. This package insert must be read completely before performing the Failure to follow the insert gives inaccurate test results.
      2. Do not open the sealed pouch unless ready to conduct the
      3. Do not use expired
      4. Bring all test materials to room temperature (15°C-30°C) before
      5. Do not use components from any other type of test kit as a substitute for the components in this
      6. Do not use hemolized blood specimens for
      7. Wear protective clothing and disposable gloves while handling the kit reagents and clinical specimens. Wash hands thoroughly after performing the
      8. Users of this test should follow the US CDC Universal Precautions for prevention of transmission of HIV, HBV and other blood-borne
      9. Dispose of all specimens and materials used to perform the test as bio-hazardous
      10. Handle the negative and positive controls in the same manner as patient
      11. The test results should be read within 15 minutes after a specimen is applied to the sample well or sample pad of the device. Reading the results after 15 minutes may give erroneous
      12. Do not perform the test in a room with strong air flow, i.e. an electric fan or strong air-conditioning.
          

          LIMITATIONS OF PROCEDURE

          • The intensity of the test line does not have a linear correlation with the antibody titer in the
          • LINEAR Anti-HCV is for screening use only. This test should be used for the detection of antibodies to HCV in serum, plasma or whole blood specimen. Any reactive specimen must be confirmed with alternative testing method(s) and clinical
          • LINEAR Anti-HCV will only indicate the presence of antibodies to HCV in the specimen and should not be used as the sole criteria for the diagnosis of Hepatitis C viral As with all diagnostic tests, all results must be considered with other clinical information available to the physician.
          • If the test result is negative and clinical symptoms persist, additional follow-up testing using other clinical methods is A negative result at any time does not preclude the possibility of Hepatitis C Virus infection. A negative result indicates absence of detectable antibodies to HCV. However, a negative test result does not preclude the possibility of exposure to or infection with HCV. A negative result can occur if the quantity of the antibodies to HCV present in the specimen is below the detection limits of the assay, or the antibodies that are detected are not present during the stage of disease in which a sample is collected.
            REFERENCES
            
            1. Kuo,G, Choo Q-L, Alter, HJ, et An assay for circulating antibodies to a major etiologic virus of human non-A, non-B hepatitis. Science 1989. 244:362-4.
            2. Esteban JI, Gonzalez A, Hernandez JM, et Evaluation of antibodies to hepatitis C virus in a study of transfusion-associated hepatitis. N Engl J Med 1990. 323:1107-12.
            3. Miyamura T, Saito I, Katayama T, et al. Detection of antibody against antigen expressed by molecularly cloned hepatitis C virus cDNA: application to diagnosis and blood screening for posttransfusion Proc Natl Acad Sci USA 1990. 87:983-7.
            4. Estaban JI, Esteban R, Viladomiu L, et Hepatitis C virus antibodies among risk groups in Spain. Lancet 1989. 2:294-7.
            5. Houghton M, Weiner A, Han J, Kuo G, Choo Q-L. Molecular Biology of the Hepatitis C viruses: Implications for diagnosis, Development, and Control of Viral Hepatology 1991. 14:381-8.
            6. Alter HJ, Purcell RH, Shih JW, Melpolder JC, Houghton M, Choo Q-L, Kuo Detection of antibody to hepatitis C virus in prospectivity followed transfusion recipients with acute and chronic non-A,non-B hepatitis. N Engl J Med 1989. 321:1494- 1500.